Sulfonamide derivatives

ABSTRACT

The present invention provides a method for inhibiting tubulin polymerization and for treating rheumatism by administering to individuals in need thereof an effective amount of sulfonamide derivatives represented by the general formula (1):                    
     wherein R 1  and R 2  may be the same or different and each independently represents a hydrogen atom, a halogen atom, a lower alkyl group, a lower alkoxy group, a nitro group, a hydroxy group; each R 3 , R 4 , R 5  and R 6  represents a hydrogen atom; A represents any group of (1) 5-membered heterocyclic group which is optionally substituted by a lower alkyl group or phenyl group, whose ring members include at least 1 nitrogen atom and may include any atom(s) selected from the group consisting of nitrogen atom, oxygen atom, and sulfur atom, (2) an alicyclic group which is optionally substituted by a lower alkyl group or phenyl group, and (3) an alicyclic group whose ring members include at least 1 nitrogen atom and may include any atom(s) selected from the group consisting of nitrogen atom, oxygen atom, and sulfur atom, or pharmaceutically acceptable salts thereof together with a pharmaceutically acceptable excipient or carrier. The sulfonamide derivatives have low toxicity and a potent effect of inhibiting tubulin polymerization.

TECHNICAL FIELD

The present invention relates to sulfonamide derivatives having tubulinpolymerization inhibitory activity, and tubulin polymerizationinhibitory agents, anticancer agents, and agents useful as preventivesor remedies for rheumatism such as inflammatory rheumatism containingthese derivatives as the active ingredients.

BACKGROUND ART

Rheumatism is a refractory disease. Rheumatoid arthritis (RA), forexample, has the basal lesion in proliferation of synovial cellsaccompanied with abnormalities in immune system caused by variousfactors. RA often causes progressive dysfunctions in articulation. Toprevent from dysfunctions by RA, which is considered to be an autoimmunedisease, agents correcting immune abnormalities are used in combinationto general antiphiogistics under the expectation of altering naturalcouse of RA.

For remedy of arthritis, steroidal agents such as adrenocorticalhormones including cortisone, non-steroidal anti-inflammatory agentssuch as aspirin, piroxicam and indometacin, antirheumatic agents such asgold preparations including aurothiomalate, D-penicillamines, andimmunosuppressive agents such as cyclophosphamide and azathioprine havebeen used.

Recent Japanese reports suggest that an intermittent administration inlow dose of methotrexate(MTX) result in high efficiency and rapidresponse. However, it has many side effects including interstitialpneumonia, stomatitis, gastrointestinal symptons such as nausea andvomition, fibrous liver, and marrow suppression. Furthermore, a longterm of the therapeutic administrations may cause high infectiousnessand complicated malignancy. There has been found no desirable drug whichis really effective and has little side effects with respect to thisdisease.

Although many compounds having tubulin polymerization inhibitoryactivity have been reported, nothing but colchicine (an arthrifuge) andvincristine (an antineoplastic) (Cancer Research, vol.20,p1023,1960)among them are applicable for therapeutic agents. Rheumacon, theglycoside extracted from the natural material having antirheumaticeffect is reported to show tubulin polymerization inhibitory effect(British Journal of rheumatology, Vol.32,p804,1993), but it has theunknown chemical structural formula and many side effects such as moonface, suffusion, and gastrointestinal disorder are reported.

The JP Laid-Open No.39256/1993 discloses that the other sulfonamidesthan those of the present invention are useful antineoplastics.N-[2-((4-hydroxyphenyl)amino)-3-pyridinyl]-4-methoxybenzenesulfonamidedescribed in the publication is reported to show tubulin polymerizationinhibitory effect (Cancer Research,Vol.54,p1702,1994).

However, the above agent has the severe side effects which makeimpossible its continuous administrations, the poor sustainedtherapeutic effects, or no effects for some patients. The clinicaltherapy demands low toxic agents which can protect and remedy patientsfrom RA through new mechanism.

DISCLOSURE OF THE INVENTION

For solving the above problem, the present inventors made diligentstudies to achieve low toxic anticancer agents and agents useful aspreventives or remedies for rheumatism such as inflammatory rheumatism.As the result, it has been found that the novel sulfonamides derivativeshaving tubulin polymerization inhibitory activity as described below arelowly toxic, effective as anticancer agents, and useful as preventivesor remedies for rheumatism such as inflammatory rheumatism. This findinghas led to the completion of the present invention.

The compounds of the present invention as shown by the generalformula(1) are novel (except the case that A is triazol in theformula(1)) and their medical uses are unknown together with the casethat A is triazol.

The present invention relates to the following (i) to (xi).

(i) Sulfonamide derivatives represented by the general formula(1)

[wherein R¹, R² may be the same or different and each independentlyrepresents a hydrogen atom, a halogen atom, a lower alkyl group, a loweralkoxy group, a nitro group, a hydroxy group, a cyano group, a C₁ to C₈acyl group, an optionally substituted phenoxy group, or an optionallysubstituted amino group;

R³, R⁴ may be the same or different and each independently represents ahydrogen atom, a halogen atom, a lower alkyl group, a lower alkoxygroup, a nitro group, a hydroxy group, a group as shown by the generalformula(2) described below:

—(CH₂)_(n)—CO—R⁷  (2)

(wherein R⁷ represents a hydrogen atom, a hydroxy group, a lower alkylgroup, a lower alkoxy group, or an optionally substituted amino group,and n means an integer of 1-5), a group as shown by the generalformula(3) described below:

—(CH₂—)_(n)—B  (3)

(wherein B represents an imidazolyl group, a triazolyl group, atetrazolyl group, or an optionally substituted amino group, and n meansany integer of 1-5), a group as shown by the general formula(4)described below:

(wherein R⁸ represents a lower alkyl group, a C₁ to C ₅ alkyl groupsubstituted by 1 or 2 hydroxy group(s), a C₁ to C₅ alkyl groupsubstituted by a C₁ to C₅ alkoxycarbonyl group, a C₁ to C₅ acyl group,or a pyridyl group; R⁹represents a lower alkyl group or a hydroxy group;a and n means any integer of 0-3 and 0-6 respectively), a group as shownby the general formula(5) described below:

(wherein R⁹ represents a lower alkyl group or a hydroxy group; b and nmeans any integer of 0-3 and 0-6 respectively), a group as shown by thegeneral formula(6) described below:

(wherein R⁹ represents a lower alkyl group or a hydroxy group; c and nmeans any integer of 0-3 and 0-6 respectively; Q represents an oxygenatom, a sulfur atom, or a group as shown by the general formula(7)described below:

(wherein d means 1 or 2)), a group as shown by the general formula(8)described below:

—(CH₂)_(n)—NR¹⁰R¹¹  (8)

(wherein R¹⁰, R₁₁ may be the same or different and each independentlyrepresents a hydrogen atom, a lower alkyl group or an optionallysubstituted amino lower alkyl group; n means any integer of 0-6), agroup as shown by the general formula(9) described below:

—(CH₂)_(n)—S(O)_(e)—R¹²  (9)

(wherein R¹² represents a hydrogen atom, a lower alkyl group or an arylgroup, an aralkyl group or an optionally substituted amino group; e andn means any integer of 0-3 and 0-6 respectively), or a group as shown bythe general formula(10) described below:

—(CH₂)_(n)—OR¹³  (10)

(wherein R¹³ represents a hydrogen atom, a lower alkyl group or an acylgroup, a phosphate group or an optionally substituted aminoalkyl group;and n means any integer of 1-6), R⁵,R⁶ may be the same or different andeach independently represents a hydrogen atom, a halogen atom, a loweralkyl group, an acyl group, or an optionally substituted amino group; Arepresents any group of (1) an optionally substituted 5-memberedheterocyclic group (except triazol) whose ring members include at least1 nitrogen atom and may include any atom(s) selected from the groupconsisting of nitrogen atom, oxygen atom, and sulfur atom, (2) anoptionally substituted alicyclic group, and (3) an alicyclic group whosering members include at least 1 nitrogen atom and may include anyatom(s) selected from the group consisting of nitrogen atom, oxygenatom, and sulfur atom], and the pharmaceutically acceptable saltsthereof.

(ii) Sulfonamide derivatives as defined in (i), wherein R¹ is a loweralkoxy group, and R², R³, R⁴, R⁵, and R⁶ may be the same or differentand each independently represents a hydrogen atom, a halogen atom, alower alkyl group, or an optionally substituted amino group, and thepharmaceutically acceptable salts thereof.

(iii) Sulfonamide derivatives as defined in (i), wherein R¹ is a loweralkoxy group, and R², R³, R⁴, R⁵, and R⁶ may be the same or differentand each independently represents a hydrogen atom, a halogen atom, alower alkyl group, or an optionally substituted amino group, and Arepresents an optionally substituted 5-membered heterocyclic group(except triazol) whose ring members include at least 1 nitrogen atom andmay include any atom(s) selected from the group consisting of nitrogenatom, oxygen atom, and sulfur atom, and the pharmaceutically acceptablesalts thereof.

(iv) Sulfonamide derivatives as defined in any one of (i)-(iii), whereinA represents an optionally substituted isoxazol group, and thepharmaceutically acceptable salts thereof.

(v) Sulfonamide derivatives as defined in (iv), wherein A represents a4-isoxazol group which has at least one lower alkyl group at the 3- and5-position, and the pharmaceutically acceptable salts thereof.

(vi) Sulfonamide derivatives as defined in (iv), wherein R¹ is a loweralkoxy group which is located at the para-position, and R² to R⁶ arehydrogen atoms, and the pharmaceutically acceptable salts thereof.

(vii) A pharmaceutical composition containing as an active ingredientthe sulfonamide derivatives (including the case that A is triazolylgroup in the general formula(1)) as defined in any one of (i)-(vi), orthe pharmaceutically acceptable salts thereof.

(vi) A tubulin polymerization inhibitory agent containing as an activeingredient the sulfonamide derivatives (including case that A istriazolyl group in the general formula(1)) as defined in any one of(i)-(vi), or the pharmaceutically acceptable salts thereof.

(ix) An anticancer agent containing as an active ingredient thesulfonamide derivatives (including the case that A is triazolyl group inthe general formula(1)) as defined in any one of (i)-(vi), or thepharmaceutically acceptable salts thereof.

(x) A drug as a preventive or a remedy for rheumatism containing as anactive ingredient the sulfonamide derivatives (including the case that Ais triazolyl group in the general formula(1)) as defined in any one of(i)-(vi), or the pharmaceutically acceptable salts thereof.

(xi) A drug as a preventive or a remedy for rheumatism as defined in(x), wherein rheumatism is inflammatory rheumatism.

BEST MODE FOR CARRYING OUT THE INVENTION

The present invention will be more particularly described below.

Lower alkyl groups of the present invention are C₁-C₆ straight chain orbranched alkyl groups including methyl group, ethyl group, n-propylgroup, isopropyl group, n-butyl group, sec-butyl group, tert-butylgroup, n-pentyl group, amyl group, isopentyl group, neopentyl group,tert-pentyl group, 1-methylbutyl group, 2-methylbutyl group,1,2-dimethylpropyl group, n-hexyl group, isohexyl group, 1-methylpentylgroup, 2-methylpentyl group, 3-methylpentyl group, 1,1-dimethylbutylgroup, 1,2-dimethylbutyl group, 2,2-dimethylbutyl group,1,3-dimethylbutyl group, 2,3-dimethylbutyl group, 3,3-dimethylbutylgroup, 1-ethylbutyl group, 2-ethylbutyl group, 1,1,2-trimethylpropylgroup, 1,2,2-trimethylpropyl group, 1-ethyl-1-methylpropyl group,1-ethyl-2-metylpropyl group. These groups may have Additionalsubstituents as long as they provide no special interference. C₁-C₄alkyl groups are preferable, C₁-C₃alkyl groups are more preferable, andmethyl group and ethyl group are the most preferable.

Lower alkoxy groups of the present invention are C₁-C₆ straight chain orbranched alkoxy groups including methoxy group, ethoxy group, isopropoxygroup, n-butoxy group, isobutoxy group, tert-butoxy group. These groupsmay have additional substituents as long as they provide no specialinterference. C₁-C₄ alkoxy groups are preferable, C₁-C₃alkoxy groups aremore preferable, and methoxy group and ethoxy group are the mostpreferable.

Halogen atoms of the present invention includes fluorine atom, chlorineatom, and bromine atom.

Substituents connected to nitrogen atoms, for example,to those found inamino groups include lower alkyl groups, and C₁-C₈ acyl groups(including C₁-C₈ alkoxycarbonyl group). These groups may have additionalsubstituents as long as they provide no special interference.

Optionally substituted amino groups include unsubstituted amino group,lower acylamino groups (for example, amino groups substituted withC₁-C₄acyl groups such as formylamino group, propionylamino group),optionally substituted alkoxycarbonylamino groups (for example,benzyloxycarbonylamino group), and mono- or di-lower alkylamino groups(for example, N,N-dimethylamino group, N,N-diethylamino group,N,N-dipropylamino group, N,N-diisopropylamino group, N,N-di-n-butylaminogroup).

Optionally substituted aminoalkyl groups include unsubstitutedaminoalkyl groups and mono- or di-lower alkyl substituted aminoalkylgroups (C₁-C₁₀) such as N,N-diethylaminoethyl group,N,N-dimethylaminomethyl group, N,N-dimethylaminoethyl group, andN,N-dimethylaminoethyl group.

Alicyclic hydrocarbon groups (groups represented by removing onehydrogen atom from the corresponding alicyclic hydrocarbon), which areincluded in A of the present invention, include 3 to 10-memberedalicyclic hydrocarbon groups such as cyclopropyl group, cyclobutylgroup, cyclopentyl group, cyclohexyl group, and cycloheptyl group. 5 to7-membered alicyclic hydrocarbon groups such as cyclohexyl group,cycloheptyl, and cycloheptyl group are preferable.

5-membered Heterocyclic hydrocarbon groups having at least one nitrogenatom, which are included in A of the present invention, mean groupsrepresented by removing one hydrogen atom from any carbon atom of thecorresponding 5-membered heterocyclic hydrocarbon rings, which have atleast two unsaturated bond and contain preferably at least twoheteroatoms. They include pyrrolyl group, oxazolyl group, isoxazolylgroup, thiazolyl group, isothiazolyl group, imidazolyl group, pyrazolylgroup, oxadiazolyl group, thiadiazolyl group, triazolyl group, andtetrazolyl group. Oxazolyl group, isoxazolyl group, and isothiazolylgroup are preferable, and isoxazolyl group is more preferable.

Alicyclic hydrocarbon groups having at least one nitrogen atom in therings, which are included in A of the present invention, mean groupsrepresented by removing one hydrogen atom from any carbon atom of thecorresponding alicyclic hydrocarbon rings, which may have oneunsaturated bond and contain preferably at least two heteroatoms. Theheteroatom may be selected from the group containing of nitrogen atom,oxygen atom, and sulfur atom. They include 3 to 10-membered alicyclichydrocarbon groups having at least one nitrogen atom in the rings suchas aziridinyl group, azethidinyl group, pyrrolidinyl group, piperidinylgroup, oxazolinyl group, isoxazolidinyl group, thiazolidinyl group,imidazolidinyl group, and pyrrazolidinyl group. Thiazolidinyl group ispreferable.

Substituents of the present invention bonding to any carbon atom ofalkyl groups or cyclic (optionally containing hetero atom(s))hydrocarbon groups include lower alkyl groups, lower alkoxy groups,optionally substituted amino groups, halogen atoms, nitro groups,hydroxy group, carboxyl groups, acyl groups (including optionallyphenyl-substituted alkoxycarbonyl), cyano group, and phenyl group. Thesegroups may have additional substituents as long as they provide nospecial interference. Lower alkyl groups are preferable for substituentsof the groups A.

Acyl groups of the present invention include C_(l)-C₈ acyl groups suchas formyl group, acetyl group, propionyl group, butyryl group, valerylgroup, optionally substituted benzoyl group, and optionally substitutedbenzyloxycarbonyl group. Substituents bonding to the benzoyl group orthe benzyloxycarbonyl group include the substituents bonding to thecarbon atom as described above.

Substituents bonding to optionally substituted phenoxy group of thepresent invention include the substituents bonding to the carbon atom asdescribed above.

Optionally substituted amino lower alkyl groups of the present inventioninclude C₁-C₃ alkylamino-substituted C₁-C₃alkyl groups, alicyclicalkylamino-substituted C₁-C₃alkyl groups, and aromatic amino-substitutedC₁-C₃alkyl groups where alicyclic alkyl groups include C₃-C₆ groups suchas cyclopropyl group, cyclobutyl group, cyclopentyl group and cyclohexylgroup, and aromatic groups include C₁-C₁₀ groups such as phenyl groupand naphthyl group.

Aralkyl groups of the present invention include C₇-C₁₁ groups such asbenzyl group.

—(CH₂)_(n)-groups of the present invention include methylene group,dimethylene group, trimethylene group, and tetramethylene group.

The groups as shown by the general formula (2) include carboxymetylgroup, methoxycarbonylmethyl group.

The groups as shown by the general formula (3) include imidazolylmethylgroup, triazolylmethyl group, tetrazolylmethyl group, aminoethyl group.

The groups as shown by the general formula(4) include4-methyl-3-methylpiperadinylmethyl group.

The groups as shown by the general formula(5) include 4-methylpiperidinyl methyl group.

The groups as shown by the general formula(6) include 1-morphorinomethyl group.

The groups as shown by the general formula(8) include dimethylaminoethylgroup.

The groups as shown by the general formula(9) include sulfonylmethylgroup.

The groups as shown by the general formula (10) include hydroxy methylgroup and methoxyethyl group.

In the present invention, preferable groups in R¹ are lower alkoxygroups located at the para position, and each preferable group in R²,R³, R⁴, R⁵, and R⁶ is hydrogen atom. Preferable groups in A includethiazolidinyl group, C₃-C₆alicyclic groups, pyrrolyl group, oxazolylgroup, isoxazolyl group. Particularly preferable in A is 4-isoxazolylgroup, which is more preferable if it has at least one substituent atthe 3- and 5-position, and is the most preferable if it has methyl groupor ethyl group both at the 3- and 5-positions. Preferable 4-isoxazolylgroups are, For example, 3-methyl-4-isoxazolyl group,3,5-dimethyl-4-isoxazolyl group, 5-ethyl-3-methyl-4-isoxazolyl group,3-ethyl-5-methyl-4-isoxazolyl group, 3,5-diethyl-4-isoxazolyl group.Combination of these preferable groups leads to the most preferablecompound among all the sulfonamide derivatives as represented by thegeneral formula(1).

Sulfonamide derivatives of the present invention may react with acids toform their salts, which are also covered by the present invention. Theirsalts include the salts of inorganic acids such as hydrogen chloride,hydrogen bromide, sulfuric acid and the salts of organic acids such asacetic acid, lactic acid, succinic acid, fumaric acid, maleic acid,citric acid, benzoic acid, methanesulfonic acid, p-toluenesulfonic acid.

The present invention covers all the hydrates and all the opticalisomers, if any.

The compounds represented by the general formula(1) include:

(1) N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

(2)N-[2-(4-methoxybenzenesulfonamide)phenyl]-1,3-dimethyl-4-(1H-pyrazole)carboxamide

(3) N-[2-(4-methoxybenzenesulfonamide)phenyl]-2-methyl-4-thiazolecarboxamide

(4) N-[2-(4-methoxybenzenesulfonamide)phenyl]-3-methyl-4-isothiazolecarboxamide

(5) N-[2-(4-methoxybenzenesulfonamide)phenyl]-2,5-dimethyl-4-oxazolecarboxamide

(6)N-[2-(4-methoxybenzenesulfonamide)phenyl]-1-methyl-1H-imidazole-4-carboxamide

(7) N-[2-(toluenesulfonamide)phenyl]-5-methyl-4-isoxazole carboxamide

(8) N-[2-(benzenesulfonamide)phenyl]-5-methyl-4-isoxazole carboxamide

(9) N-[2-(4-fluorobenzenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

(10) N-[2-(4-nitrobenzenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

(11) N-[2-(3,4-dimethoxybenzenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

(12) N-[2-(4-methoxybenzenesulfonamide)phenyl]-(L)-prolinamide

(13) N-[2-(4-methoxybenzenesulfonamide)phenyl]-3,5-dimethyl-4-isoxazolecarboxamide

(14) N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-ethyl-4-isoxazolecarboxamide

(15) (±)-N-[2-(4-methoxybenzenesulfonamide)phenyl]-4-thiazolidinecarboxamide

(16)(±)-N-[2-(4-methoxybenzenesulfonamide)phenyl]-3-(N-methylpiperidine)carboxamide

(17) N-[2-(4-methoxybenzenesulfonamide)phenyl]-(D)-prolinamide

(18) N-[2-(4-methoxybenzenesulfonamide)phenyl]-4-piperidine carboxamide

(19) (±)-N-[2-(4-methoxybenzenesulfonamide)phenyl]-3-piperidinecarboxamide

(20) (±)-N-[2-(4-methoxybenzenesulfonamide)phenyl]-2-piperidinecarboxamide

(21) N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-methyl-4-oxazolecarboxamide

(22) N-[2-(4-methoxybenzenesulfonamide)phenyl]-cyclohexyl carboxamide

(23) N-[2-(4-methoxybenzenesulfonamide)phenyl]-cyclopropyl carboxamide

(24) N-[2-(4-methoxybenzenesulfonamide)phenyl]-cyclobutyl carboxamide

(25) N-[2-(4-methoxybenzenesulfonamide)phenyl]-cyclopentyl carboxamide

(26) N-[2-(4-methoxybenzenesulfonamide)phenyl]-N-methyl-2-pyrrolecarboxamide

(27)N-[2-(4-methoxybenzenesulfonamide)phenyl]-3-ethyl-5-methyl-4-isoxazolecarboxamide

(28)N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-ethyl-3-methyl-4-isoxazolecarboxamide

(29) N-[2-(4-methoxybenzenesulfonamide)phenyl]-3,5-diethyl-4-isoxazolecarboxamide

(30)N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-isopropyl-3-methyl-4-isoxazolecarboxamide

(31)N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-methyl-3-phenyl-4-isoxazolecarboxamide

(32)N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-methyl-3-phenyl-4-isoxazolecarboxamide

The compounds of the present invention, which are novel and have not yetpublished in any documents, can be produced, for example, by thefollowing reaction:

The novel sulfonamide derivatives represented by the general formula(1)can be produced by reacting compounds as shown by general formula(11)described below,

(wherein R¹, R², R³, R⁴, R⁵, and R⁶ have the same meaning as mentionedbefore) with carboxylic acids or their reactive derivatives (whosefunctional groups may be protected if they are not associated with thereaction) as shown by general formula(12) described below,

A—Z  (12)

(A has the same meaning as mentioned before, and Z represents carboxylgroups or the reactive groups derived therefrom).

The reaction is preferably carried out under the presence of a base.

Most of the compounds represented by the general formula(11) are knownto the public, and the others can be prepared by the method JP Laid-OpenNo.39256/1993 has disclosed, or its modifications.

Any reactive derivatives that are generally used to form carboxamidebonds can be applied for reactive derivatives of carboxylic acidsrepresented by the general formula(12). They include acid halides,active amides, and active esters. They may be prepared prior to thereaction or planned to form within the reaction system.

The acid halides, which include acid chloride and acid bromide, can beobtained generally by reacting carboxylic acids with thionylhalogenides.

For the active amides, there can be used acid amides of, for example,imidazole, pyrazole, 4-substituted imidazole, dimethylpyrazole,triazole, tetrazole, and benzothiazole.

For the active esters, there can be used acid esters which includemethyl esters, methoxymethyl esters, cyanomethyl esters, propagylesters, 4-nitrophenyl esters, 2,4-dinitrophenyl esters, and esters of,for example, 1-hydroxybenzotriazole.

The carboxylic acids represented by the general formusa(12) may bereacted with the amines represented by the general formula(11) under thepresence of condensation agents such asN,N′-dicyclohexylcarbodiimide(DCC) and N-cyclohexyl-N-morpholinoetylcarbodiimide. These reactions are preferably carried out under thepresence of bases such as organic tertiary amines (for example,triethylamine, N,N-dimethylaniline, and pyridine).

The reaction solvents, which are desirable to use if they can dissolvethe reaction-associated materials and have no reaction with them,include pyridine, tetrahydrofurane, dioxane, benzene, ether, methylenechloride, dimethylformamide, toluene, and the mixture solventsconsisting of two or more solvents selected therefrom. But any reactionsolvents can be used with no special limitation regardless of the abovementioned examples.

The reaction can be generally carried out at ambient temperature.Cooling or heating may be applied for the reaction if necessary. Thereaction time is generally 5 min-20 hrs, and may be determinedadequately depending upon materials and reaction temperature.

Deprotection such as acid treatment converts the reaction products,whose amino groups are protected, to the compounds represented by thegeneral formula(1) which have the free amino groups.

The representative compounds as shown by the general formula(11) whichare raw materials for syntheses include:

N-(2-aminophenyl)-4-methoxybenzenesulfonamide

N-(2-aminophenyl)-4-ethoxybenzenesulfonamide

N-(2-aminophenyl)-4-propoxybenzenesulfonamide

N-(2-aminophenyl)-3,4-dimethoxybenzenesulfonamide

N-(2-aminophenyl)-4-toluenesulfonamide

N-(2-aminophenyl)-4-fluorobenzenesulfonamide

N-(2-aminophenyl)-4-nitrobenzenesulfonamide

The representative compounds as shown by the general formula(12) whichare raw materials for syntheses include:

5-methylisoxazole-4-carboxylic acid

5-ethylisoxazole-4-carboxylic acid

3-ethyl-5-methylisoxazole-4-carboxylic acid

5-ethyl-3-methylisoxazole-4-carboxylic acid

3,5-diethylisoxazole-4-carboxylic acid

3,5-dimethylisoxazole-4-carboxylic acid

1,3-dimethyl-1H-pyrazole-4-carboxylic acid

2-methylthiazole-4-carboxylic acid

2,5-dimethyloxazole-4-carboxylic acid

N-methyl-1H-imidazole-4-carboxylic acid

N-(t-butoxycarbonyl)-L-proline

N-(t-butoxycarbonyl)-D-proline

N-(t-butoxycarbonyl)thiazolidine-4-carboxylic acid

N-methylpiperidine-3-carboxylic acid

N-(t-butoxycarbonyl)piperidine-4-carboxylic acid

N-(t-butoxycarbonyl)piperidine-3-carboxylic acid

N-(t-butoxycarbonyl)piperidine-2-carboxylic acid

cyclopropane carboxylic acid

cyclobutane carboxylic acid

cyclopentane carboxylic acid

cyclohexane carboxylic acid

1-metyl-2-pyrrole carboxylic acid

The sulfonamide derivatives represented by the general formula(1) or thepharmaceutically acceptable salts thereof may be administrated orally orparenterally (systemic or local effect) for medical use either alone orin various preparation forms such as pulver, granule, tablet, andinjection manufactured by mixing with pharmaceutically acceptableadditives such as vehicle, excipient, diluent, and solubilizer.Preparations should contain 0.1-100% by weight of the compounds of thepresent invention or the pharmaceutically acceptable salts thereofdepending on the preparation forms. The doses should be determineddepending on administration routes, patients' ages, and disease symptomsto protect or cure from. The dose, for example, by oral administrationfor an adult is 0.1 mg-2,000 mg/day, and is preferably 1 mg-1,000 mg/dayin one or more times per day.

The sulfonamide derivatives represented by the general formula(1) andthe pharmaceutically acceptable salts thereof have tubulinpolymerization inhibitory activity, and are useful as anticancer agentsand preventives or remedies for rheumatism. Rheumatism herein includes,for example, inflammatory rheumatism such as rheumatoid artiritis andosteoarthritis etc.

EXAMPLE

The present invention will now be described more in details by way ofexamples, provided that the present invention should not be limited bythese examples. The efficiency of the compounds according to the presentinvention will be demonstrated by way of test examples with respect topharmacological test results of the representative compounds. NMR valueswere measured by 200 Mz NMR with tetramethylsilane set as an internalstandard.

Example 1 N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

To the suspension of 5-methylisoxazole-4-carboxylic acid (2.70 g, 21.3mmol) in methylene chloride (10 ml) was added pyridine (3.36 g, 42.5mmol) under ice-cooled nitrogen atmosphere before thionyl chloride (2.53g, 21.3 mmol) was added to dropwise, and then the solution was stirredat the same temperature for 30 min. To the resultant solution was addedthe solution of N-(2-aminophenyl)-4-methoxybenzene sulfonamide(JPLaid-Open No.39256/1993) (4.93 g, 17.7 mmol) in methylene chloride(20 ml), and the solution was left to return back to ambient temperaturegradually before it was stirred overnight. The reaction solution wasadded to aqueous saturated sodium bicarbonate, and then subject toextraction by chloroform. The organic layer was dried over anhydroussodium sulfate, and then concentrated to obtain the crude crystal. Itwas purified by recrystallization from ethanol to yield 4.50 g of thetitled compound.

NMR (CDCl₃) ppm: 2.80 (3H, s), 3.85 (3H, s) 6.53-6.59 (2H, m), 6.89 (2H,d, J=9.0 Hz), 6.94 (1H, m), 7.28 (1H, dt, J=1.5, 7.6 Hz), 7.59 (2H, d,J=9.0 Hz), 8.08 (1H, dd, J=1.4, 8.2 Hz), 8.20 (1H, s), 8.96 (1H, br s).

MS (FAB, POS) m/z: 388 [M+H]⁺.

Example 2N-[2-(4-methoxybenzenesulfonamide)phenyl]-1,3-dimethyl-4-(1H-pyrazole)carboxamide

Like the Example 1 process, 1,3-dimethyl-1H-pyrazole-4-carboxylic acid(1.54 g, 11.0 mmol) was made to react with N-(2-aminophenyl)-4-methoxybenzene sulfon amide (2.70 g, 10.0 mmol) to yield 1.54 g of the titledcompound.

NMR (CDCl₃) ppm: 2.54 (3H, s), 3.84 (3H, s), 3.87 (3H, s), 6.57 (1H, dd,J=1.4, 7.9 Hz), 6.81-6.92 (3H, d+m, J=9.0 Hz), 6.97 (1H, br s), 7.20(1H, m), 7.59 (2H, d, J=9.0 Hz), 7.85 (1H, s), 8.20 (1H, dd, J=1.4, 8.3Hz), 8.65 (1H, br s).

MS (FAB, POS) m/z: 401 [M+H]⁺.

Example 3 N-[2-(4-methoxybenzenesulfonamide)phenyl]-2-methyl-4-thiazolecarboxamide

To the suspension of 5-methylthiazole-4-carboxylic acid (510 mg, 3.6mmol) in methylene chloride (3.6 ml) was added pyridine (295 mg, 3.6mmol) at ambient temperature before thionyl chloride (473 mg, 4.0 mmol)was added to dropwise. After 30 min, the resultant solution was added tothe solution of N-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.00g,3.6 mmol) in methylene chloride (7.2 ml) with pyridine (378 mg, 3.6mmol) further added thereto, before it was stirred overnight. To thereaction solution was added water and ethyl acetate to extract. Theorganic layer was washed with aqueous saturated sodium bicarbonate,dried over anhydrous magnesium sulfate, and then concentrated to obtainthe crude crystal. It was washed with diethyl ether and purified byrecrystallization from isopropyl alcohol to yield 390 mg of the titledcompound.

NMR (DMSO-d₆) ppm: 2.79 (3H, s), 3.77 (3H, s), 6.84 (1H, dd, J=1.5, 7.9Hz), 6.95 (2H, d, J=9.0 Hz), 7.03 (1H, dt, J=1.5, 7.9 Hz), 7.28 (1H, dt,J=1.5, 8.2 Hz), 7.58 (2H, d, J=9.0 Hz), 8.13 (1H, dd, J=1.5, 8.2 Hz),8.25 (1H, s), 9.68 (1H, br s), 10.00 (1H, br s).

MS (FAB, POS) m/z: 404 [M+H]⁺.

Example 4N-[2-(4-methoxybenzenesulfonamide)phenyl]-3-methyl-4-isothiazolecarboxamide

Like the Example 3 process, 3-methylisothiazole-4-carboxylic acid (510mg, 3.7 mmol) was made to react with N-(2-aminophenyl)-4-methoxybenzenesulfon amide (1.00 g, 3.6 mmol) to yield 390 mg of the titled compound.

NMR (DMSO-d₆) ppm: 2.56 (3H, s), 3.75 (3H, s), 6.91 (2H, d, J=9.0 Hz),7.11-7.23 (3H, m), 7.53 (2H, d, J=9.0 Hz), 7.66 (1H, d, J=7.6 Hz), 9.41(1H, br s), 9.42 (1H, s), 9.47 (1H, br s).

MS (FAB, POS) m/z: 404 [M+H]⁺.

Example 5N-[2-(4-methoxybenzenesulfonamide)phenyl]-2,5-dimethyl-4-oxazolecarboxamide

Like the Example 1 process, 2,5-dimethyloxazole-4-carboxylic acid (1.40g, 9.9 mmol) was made to react with N-(2-aminophenyl)-4-methoxybenzenesulfonamide (2.50 g, 9.0 mmol) to yield 1.90 g of the titled compound.

NMR (CDCl₃) ppm: 2.45 (3H, s), 2.64 (3H, s), 3.80 (3H, s), 6.76 (2H, d,J=9.0 Hz), 7.11-7.42 (4H, m), 7.56 (2H, d, J=9.0 Hz), 7.76 (1H, br s),8.62 (1H, br s).

MS (FAB, POS) m/z: 402 [M+H]⁺.

Example 6N-[2-(4-methoxybenzenesulfonamide)phenyl]-1-methyl-1H-imidazole-4-carboxamide

To the suspension of N-methyl-1H-imidazole-4-carboxylic acid (594 mg,3.3 mmol) in methylene chloride (3 ml) was added pyridine (569 mg, 7.2mmol) under ice-cooled nitrogen atmosphere before thionyl chloride (428mg, 3.6 mmol) was added to dropwise, and then the solution was stirredat the same temperature for 30 min. To the resultant solution was addedthe solution of N-(2-aminophenyl)-4-methoxy benzenesulfonamide (834 mg,3.0 mmol) in methylene chloride (3 ml), and the solution was left toreturn back to ambient temperature gradually before it was stirredovernight. The reaction solution was added to aqueous saturated sodiumbicarbonate, and then subject to extraction by chloroform. The organiclayer was dried over anhydrous sodium sulfate, and then concentrated toobtain the residue. It was purified by silicagel chromatography(chloroform:methanol=19:1) to yield 100 mg of the titled compound.

NMR (DMSO-d₆) ppm: 3.74 (3H, d, J=4.2 Hz), 3.80 (3H, s), 6.67 (1H, dd,J=1.3, 7.9 Hz), 6.88-7.09 (3H, d+m, J=8.9 Hz), 7.18-7.37 (2H, m), 7.61(2H, d, J=8.9 Hz), 7.75-7.85 (3H, m), 8.22 (1H, dd, J=1.3, 8.2 Hz), 9.63(1H, br s), 9.88 (1H br s).

MS (FAB, POS) m/z: 387 [M+H]⁺.

Example 7 N-[2-(p-toluenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

Like the Example 1 process, 5-methylisoxazole-4-carboxylic acid (667 mg,5.3 mmol) was made to react with N-(2-aminophenyl )-4-toluenesulfonamide (1.31 g, 5.0 mmol) to yield 1.19 g of the titled compound.

NMR (DMSO-d₆) ppm: 2.29 (3H, s), 2.67 (3H, s), 7.13-7.25 (5H, d+m, J=8.3Hz), 7.45-7.55 (3H, d+m, J=8.3 Hz), 8.82 (1H, s), 9.30 (1H, s), 9.44(1H, s).

MS (FAB, POS) m/z: 372[M+H]⁺.

Example 8 N-[2-(benzenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

Like the Example 1 process, 5-methylisoxazole-4-carboxylic acid (667 mg,5.3 mmol) was made to react with N-(2-aminophenyl)-benzene sulfonamide(1.23 g, 5.0 mmol) to yield 690 mg of the titled compound.

NMR (DMSO-d₆) ppm: 2.66 (3H, s), 7.14-7.26 (3H, m), 7.37-7.65 (6H, m),8.84 (1H, s), 9.32 (1H, s), 9.56 (1H,

MS (FAB, POS) m/z: 358 [M+H]⁺.

Example 9 N-[2-(4-fluorobenzenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

Like the Example 1 process, 5-methylisoxazole-4-carboxylic acid (667 mg,5.3 mmol) was made to react with N-(2-aminophenyl)-4-fluorobenzenesulfonamide (1.33 g, 5.0 mmol) to yield 300 mg of the titled compound.

NMR (DMSO-d₆) ppm: 2.66 (3H, s), 7.16-7.28 (5H, m), 7.49 (1H, m),7.59-7.69 (2H, m), 8.85 (1H, s), 9.31 (1H, br s), 9.56 (1H, br s).

MS (FAB, POS) m/z: 376 [M+H]⁺.

Example 10 N-[2-(4-nitrobenzenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

Like the Example 1 process, 5-methylisoxazole-4-carboxylic acid (667 mg,5.3 mmol) was made to react with N-(2-aminophenyl)-4-nitrobenzenesulfonamide (1.46 g, 5.0 mmol) to yield 330 mg of the titled compound.

NMR (DMSO-d₆) ppm: 2.59 (3H, s), 7.20-7.33 (3H, m), 7.43 (1H, m), 7.81(2H, d, J=8.8 Hz), 8.21 (2H, d, J=8.8 Hz), 8.82 (1H, s), 9.27 (1H, brs), 9.80 (1H, br s).

MS (FAB, POS) m/z: 403 [M+H]⁺.

Example 11N-[2-(3,4-dimethoxybenzenesulfonamide)phenyl]-5-methyl-4-isoxazolecarboxamide

Like the Example 6 process, 5-methylisoxazole-4-carboxylic acid (667 mg,5.3 mmol) was made to react with N-(2-aminophenyl)-3,4-dimethoxybenzenesulfonamide (1.54 g, 5.0 mmol) to yield 750 mg of the titled compound.

NMR (DMSO-d₆) ppm: 2.63 (3H, s), 3.56 (3H, s), 3.75 (3H, s), 6.89 (1H,d, J=8.6 Hz), 6.97 (1H, d, J=2.2 Hz), 7.10-7.31 (5H, m), 7.50 (1H, m),8.80 (1H, s), 9.23 (1H, s), 9.32 (1H, s).

MS (FAB, POS) m/z: 418 [M+H]⁺.

Example 12 N-[2-(4-methoxybenzenesulfonamide)phenyl]-(L)-prolinamide HClsalt

To the suspension of N-(t-butoxycarbonyl)-L-prolin (474 mg, 2.2 mmol) inmethylene chloride (5 ml) was added pyridine (348 mg, 4.4 mmol) underice-cooled nitrogen atmosphere before thionyl chloride (367 mg, 2.2mmol) was added to dropwise, and then the solution was stirred at thesame temperature for 30 min. To the resultant solution was added thesolution of N-(2-aminophenyl)-4-methoxy benzenesulfonamide (556 mg, 2.0mmol), and the solution was left to return back to ambient temperaturegradually before it was stirred overnight. The reaction solution wasconcentrated to obtain the residue, to which 4N-HCl/dioxane (5.0 ml,20.0mmol) was added before it was stirred at ambient temperature for 2 hrs.The reaction solution was concentrated to obtain the residue, which waspurified by silicagel chromatography (chloroform:methanol=19:1) andHP-20 (0 to 90% aqueous methanol solution) to yield 210 mg of the titledcompound.

NMR (CD₃OD) ppm: 2.05-2.22 (2H, m), 2.31 (1H, m), 2.55 (1H, m),3.34-3.55 (2H, m), 3.86 (3H, s), 4.55 (1H, t, J=8.0 Hz), 6.70 (1H, d,J=7.9 Hz), 7.00 (2H, d, J=8.9 Hz), 7.02 (1H, m), 7.25 (1H, m), 7.58 (2H,d, J=8.9 Hz), 7.78 (1H, m).

MS (FAB, POS) m/z: 376 [M+H]⁺.

Example 13N-[2-(4-methoxybenzenesulfonamide)phenyl]-3,5-dimethyl-4-isoxazolecarboxamide

Like the Example 6 process, 3,5-dimethylisoxazole-4-carboxylic acid (846mg, 6.0 mmol) was made to react with N-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.39 g, 5.0 mmol) to yield 600 mg of the titled compound.

NMR (DMSO-d₆) ppm: 2.41 (3H, s), 2.63 (3H, s), 3.82 (3H, s), 6.83 (1H,dd, J=1.4, 8.0 Hz), 6.99-7.09 (3H, m), 7.22 (1H, dt, J=1.5, 8.0 Hz),7.60 (2H, d, J=9.0 Hz), 7.86 (s), 9.10 (1H, brs), 9.49 (1H, brs),

MS (FAB, POS) m/z: 402 [M+H]⁺.

Example 14 N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-ethyl-4-isoxazolecarboxamide

Like the Example 1 process, 5-ethylisoxazole-4-carboxylic acid (846 mg,6.0 mmol) was made to react with N-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.39 g, 5.0 mmol) to yield 1.04 g of the titled compound.

NMR (CDCl₃) ppm: 1.39 (3H, t, J=7.6 Hz), 3.23(2H, q, J=7.6 Hz), 3.85(3H, s), 6.58 (1H, dd, J=1.4, 8.0 Hz), 6.63 (1H, s), 6.89 (2H, d, J=9.0Hz), 6.94 (1H, dt, J=1.5, 7.7 Hz), 7.28 (1H, m), 7.58 (2H, d, J=9.0 Hz),8.07 (1H, dd, J=1.4, 8.0 Hz), 8.60 (1H, br s).

MS (FAB, POS) m/z: 402 [M+H]⁺.

Example 15 (±)-N-[2-(4-methoxybenzenesulfonamide)phenyl]-4-thiazolidinecarboxamide HCl salt

To the suspension of N-(t-butoxycarbonyl)thiazolidine-4-carboxylic acid(1.0 g, 6.0 mmol) in methylene chloride (10 ml) was added pyridine (1.02g, 12.0 mmol) under ice-cooled nitrogen atmosphere before thionylchloride (718 mg, 6.0 mmol) was added to dropwise, and then the solutionwas stirred at the same temperature for 30 min. To the resultantsolution was added the solution of N-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.39 g, 5.0 mmol), and the solution was left toreturn back to ambient temperature gradually before it was stirredovernight. The reaction solution was concentrated to obtain the residue,to which ethyl acetate and aqueous saturated sodium bicarbonate wereadded to extract. The organic layer was dried over anhydrous sodiumsulfate and concentrated to obtain the residue, which was purified bysilicagel chromatography (chloroform:methanol=19:1). To the resultantresidue was added 4N-HCl/dioxane (12.5 ml, 50.0 mmol) before it wasstirred at ambient temperature for 2 hrs. The reaction solution wasconcentrated to obtain the crude crystal, which was purified byrecrystallization from methanol to yield 1.05 g of the titled compound.

NMR (CD₃OD) ppm: 3.11 (1H, dd, J=7.5, 10.7 Hz), 3.40 (1H, dd, J=4.0,10.7 Hz), 3.86 (3H, s), 4.16 (1H, d, J=9.7 Hz), 4.32 (1H, d, J=9.7 Hz),4.33 (1H, dd, J=2.0, 7.5 Hz), 6.64 (1H, dd, J=1.5, 7.0 Hz), 6.90-7.04(3H, m+d, J=9.0 Hz), 7.24 (1H, dt, J=1.5, 8.0 Hz), 7.59 (2H, d, J=9.0Hz), 8.02 (1H, dd, J=1.4, 8.0 Hz).

MS (FAB, POS) m/z: 394 [M+H]⁺.

Example 16 (±)-N-[2-(4-methoxybenzenesulfonamide)phenyl]-3-(N-methylpiperidine) carboxamide HCl salt

To the suspension of N-methylpiperidine-3-carboxylic acid (537 mg, 3.0mmol) in methylene chloride (10 ml) was added pyridine (767 mg, 9.0mmol) under ice-cooled nitrogen atmosphere before thionyl chloride (359mg, 3.0 mmol) was added to dropwise, and then the solution was stirredat the same temperature for 30 min. To the resultant solution was addedthe solution of N-(2-aminophenyl)-4-methoxy benzenesulfonamide (695 mg,2.5 mmol), and the solution was left to return back to ambienttemperature gradually before it was stirred overnight. The reactionsolution was concentrated to obtain the residue, which was purified byHP-20 (gradient with 0 to 90% aqueous mathanol solution) to yield 550 mgof the titled compound.

NMR (DMSO-d₆) ppm: 1.42 (1H, m), 1.80-2.02 (3H, m), 2.79 (3H, d, J=3.4Hz), 2.85-3.50 (5H, m), 3.82 (3H s), 7.01-7.20 (5H, d+m, J=9.0 Hz),7.51-7.59 (3H, m+d, J=9.0 Hz), 9.45 (1H, br s), 9.65 (1H, br s).

MS (FAB, POS) m/z: 440 [M+H]⁺.

Example 17 N-[2-(4-methoxybenzenesulfonamide)phenyl]-(D)-prolinamide HClsalt:

Like the Example 12 process, N-(t-butoxycarbonyl)-D-prolin (1.29 g, 6.0mmol) was made to react with N-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.39 g, 5.0 mmol) to yield 150 mg of the titled compound.

NMR (CD₃OD) ppm: 1.90-2.08 (2H, m), 2.16 (1H, m) 2.42 (1H, m), 3.20-3.36(2H, m), 3.87 (3H, s), 4.23 (1H, d d, J=6.0, 8.9 Hz), 6.68 (1H, dd,J=1.5, 8.0 Hz), 6.94-7.03 (3H, d+m, J=9.0 Hz), 7.25 (1H, dt, J=1.5, 8.0Hz), 7.59 (2H, d, J=9.0 Hz), 7.90 (1H, dd, J=1.4, 8.2 Hz).

MS (FAB, POS) m/z: 376 [M+H]⁺.

Example 18 N-[2-(4-methoxybenzenesulfonamide)phenyl]-4-piperidinecarboxamide HCl salt:

To the suspension of N-(t-butoxycarbonyl)piperidine-4-carboxylic acid(1.38 g, 6.0 mmol) in methylene chloride (20 ml) was added pyridine(1.02 g, 12.0 mmol) under ice-cooled nitrogen atmosphere before thionylchloride (718 mg, 6.0 mmol) was added to dropwise, and then the solutionwas stirred at the same temperature for 30 min. To the resultantsolution was added the solution of N-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.39 g, 5.0 mmol), and the solution was left toreturn back to ambient temperature gradually before it was stirredovernight. The reaction solution was concentrated to obtain the residue,to which ethyl acetate and aqueous saturated sodium bicarbonate wereadded to extract. The organic layer was dried over anhydrous sodiumsulfate and concentrated to obtain the residue, to which 4N-HCl/dioxane(15.0 ml,60.0 mmol) was added before it was stirred at ambienttemperature for 2 hrs. The reaction solution was concentrated to obtainthe crude crystal, which was washed with isopropyl alcohol to yield 1.52g of the titled compound.

NMR (DMSO-d₆) ppm: 1.67-2.00 (4H, m), 2.61 (1H, m), 2.83-3.06 (2H, m)3.25-3.44 (3H, m), 3.80 (3H, s), 6.9 7-7.20 (5H, d+m, J=8.9 Hz), 7.55(2H, d, J=8.9 Hz), 7.64 (1H, d, J=7.7 Hz), 8.65 (1H. br s), 8.90 (1H, brs).

MS (FAB, POS) m/z: 390 [M+H]⁺.

Example 19 (±)-N-[2-(4-methoxybenzenesulfonamide)phenyl]-3-piperidinecarboxamide HCl salt

Like the Example 18 process, N-(t-butoxycarbonyl)piperidine-3-carboxylicacid (1.38 g, 6.0 mmol) was made to react withN-(2-aminophenyl)-4-methoxybenzene sulfonamide (1.39 g, 5.0 mmol) toyield 900 mg of the titled compound, where the corresponding residue waspurified by silicagel chromatography (chloroform:methanol=19:1) andHP-20 (gradient with 0 to 90% aqueous methanol solution).

NMR (CD₃OD) ppm: 1.82-2.21 (5H, m), 2.91-3.48 (4H, m), 3.86 (3H, s),6.84 (1H, dd, J=1.5, 7.9 Hz), 6.98 (2H, d, J=8.9 Hz), 7.04 (1H, dt,J=1.5, 7.9 Hz), 7.23 (1H, dt, J=1.4, 8.0 Hz), 7.56 (2H, d, J=8.9 Hz),7.69 (1H, dd, J=1.4, 8.0 Hz).

MS (FAB, POS) m/z: 390 [M+H]⁺.

Example 20 (±)-N-[2-(4-methoxybenzenesulfonamide)phenyl]-2-piperidinecarboxamide HCl salt

Like the Example 18 process, N-(t-butoxycarbonyl)piperidine-3-carboxylicacid (1.38 g, 6.0 mmol) was made to react withN-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.39 g,5.0 mmol) to yield1.24 g of the titled compound, where the corresponding residue waspurified by HP-20 (gradient with 0 to 90% aqueous methanol solution).

NMR (CD₃OD) ppm: 1.65-2.07 (5H, m), 2.42 (1H, m), 3.09 (1H, m), 3.45(1H, m), 3.85 (3H, s), 4.10 (1H, m), 6.70 (1H, dd, J=1.5, 7.9 Hz), 6.99(2H, d, J=9.0 Hz), 7.02 (1H, m), 7.25 (1H, dt, J=1.5, 8.0 Hz), 7.59 (2H,d, J=9.0 Hz), 7.83 (1H, dt, J=1.4, 8.0 Hz).

MS (FAB, POS) m/z: 390 [M+H⁺.

Example 21 N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-methyl-4-oxazolecarboxamide

Like the Example 1 process, 5-methyloxazole-4-carboxylic acid (315 mg,2.5 mmol) was made to react with N-(2-aminophenyl)-4-methoxybenzenesulfonamide (584 mg, 2.1 mmol) to yield 430 mg of the titled compound.

NMR (DMSO-d₆) ppm: 2.64 (3H, s), 3.80 (3H, s), 6.76 (1H, dd, J=1.4, 7.9Hz), 6.94 (2H, d, J=9.0 Hz), 6.98 (1H, dt, J=1.5, 7.9 Hz), 7.24 (1H, dt,J=1.5, 8.2 Hz), 7.56 (2H, d, J=9.0 Hz), 8.11 (1H, dd, J=1.4, 8.2 Hz),8.42 (1H, s), 9.63 (1H, br s), 9.75 (1H, br s).

MS (FAB, POS) m/z: 388 [M+H]⁺.

Example 22 N-[2-(4-methoxybenzenesulfonamide)phenyl]-cyclohexylcarboxamide

To the solution of N-(2-aminophenyl)-4-methoxybenzene sulfon amide (1.39g, 5.0 mmol) in tetrahydrofuran (20 ml) was dropwise added cyclohexanecarbonyl chloride (748 mg, 5.1 mmol) under ice-cooled nitrogenatmosphere, and then the solution was left to return back to ambienttemperature gradually before it was stirred overnight. The reactionsolution was concentrated to obtain the residue, which was dissolved inethyl acetate and washed with aqueous sodium bicarbonate. The organiclayer was dried over anhydrous sodium sulfate, and concentrated toobtain the crude crystal, which was purified by recrystallization fromethanol to yield 1.10 g of the titled compound.

NMR (DMSO-d₆) ppm: 1.16-1.43 (5H, m), 1.60-1.83 (5H, m), 2.16 (1H, m),3.81 (3H, s), 6.95-7.20 (5H, m+d, J=8.8 Hz), 7.46-7.67 (3H, m+d, J=8.8Hz), 9.02 (1H, s), 9.23 (1H, s).

MS (FAB, POS) m/z: 389 [M+H]⁺.

Example 23 N-[2-(4-methoxybenzenesulfonamide)phenyl]-cyclopropylcarboxamid

Like the Example 22 process, N-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.39 g, 5.0 mmol) was made to react with cyclopropanecarbonyl chloride (533 mg, 5.0 mmol) to yield 820 mg of the titledcompound.

NMR (DMSO-d₆) ppm: 0.75-0.86 (4H, m), 1.60 (1H, m), 3.80 (3H, s), 6.99(2H, d, J=9.0), 7.05-7.20 (3H, m), 7.45-7.54 (3H, m), 9.20 (1H, br s),9.45 (1H, br s).

MS (FAB, POS) m/z: 347 [M+H]⁺.

Example 24 N-[2-(4-methoxybenzenesulfonamide)phenyl]-cyclobutylcarboxamid

Like the Example 22 process, N-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.39 g, 5.0 mmol) was made to react with cyclobutanecarbonyl chloride (597 mg, 5.1 mmol) to yield 1.54 g of the titledcompound.

NMR (DMSO-d₆) ppm: 1.75-2.25 (6H, m), 3.10 (1H, m), 3.79 (3H, s),6.97-7.22 (5H, m), 7.43-7.60 (3H, m) 9.00 (1H, br s), 9.28 (1H, br s).

MS (FAB, POS) m/z: 361 [M+H]⁺.

Example 25 N-[2-(4-methoxybenzenesulfonamide)phenyl]-cyclopentylcarboxamid

Like the Example 22 process, N-(2-aminophenyl)-4-methoxybenzenesulfonamide (1.39 g, 5.0 mmol) was made to react with cyclopentanecarbonyl chloride (666 mg, 5.0 mmol) to yield 1.02 g of the titledcompound.

NMR (DMSO-d₆) ppm: 1.50-1.90 (8H, m), 2.65 (1H, m), 3.80 (3H, s),6.97-7.22 (5H, m), 7.46-7.62 (3H, m), 9.15 (1H, br s), 9.26 (1H, br s).

MS (FAB, POS) m/z: 375 [M+H]⁺.

Example 26 N-[2-(4-methoxybenzenesulfonamide)phenyl]-N-methyl-2-pyrrolecarboxamide

Like the Example 1 process, 1-methyl-2-pyrrole carboxylic acid (625 mg,5.0 mmol) was made to react with N-(2-aminophenyl)-4-methoxybenzenesulfon amide (1.39 g, 5.0 mmol) to yield 1.55 g of the titled compound.

NMR (DMSO-d₆) ppm: 3.77 (3H, s), 3.84 (3H, s), 6.14 (1H, dd, J=2.6, 4.0Hz), 6.84 (1H, dd, J=2.0, 4.0 Hz), 6.93 (2H, d, J=9.0 Hz), 6.97-7.09(3H, m), 7.22 (1H, dt, J=2.0, 8.1 Hz), 7.57 (2H, d, J=9.0 Hz), 7.76 (1H,dd, J=1.3, 8.0 Hz), 9.22 (1H, br s), 9.54 (1H, br s).

MS (FAB, POS) m/z: 386 [M+H]⁺.

Example 27N-[2-(4-methoxybenzenesulfonamide)phenyl]-3-ethyl-5-methyl-4-isoxazolecarboxamide

To the suspension of 3-ethyl-5-methylisoxazole-4-carboxylic acid (930mg, 6.0 mmol) in methylene chloride (10 ml) was added pyridine (0.97 ml,12.0 mmol) under ice-cooling by stirring before thionyl chloride (0.44ml, 6.0 mmol) was added to dropwise. After 30 min, to the resultantsolution was added N-(2-aminophenyl)-4-methoxy benzenesulfonamide (1.39g, 5.0 mmol) under ice-cooling, and the solution was left to return backto ambient temperature gradually before it was stirred overnight. To thereaction solution was added water and ethyl acetate to extract. Theorganic layer was washed with aqueous saturated sodium bicarbonate,2N-HCl, and water, and then dried over anhydrous magnesium sulfate. Theresultant solution was concentrated to obtain the crude residue, whichwas purified by silicagel chromatography (hexane:ethyl acetate=2:1) toyield 1.95 g of the titled compound.

NMR (DMSO-d₆) ppm: 1.24 (3H, t, J=7.5 Hz), 2.63 (3H, s), 2.84 (2H, q,J=7.5 Hz), 3.83 (3H, s), 6.84 (1H, dd, J=1.5, 7.9 Hz), 6.95-7.08 (3H,m), 7.21 (1H, dt, J=1.5, 7.9 Hz), 7.60 (2H, d, J=8.8 Hz), 7.86 (1H, dd,J=1.3, 8.1 Hz), 9.10 (1H, br s), 9.45 (1H, br s).

MS (FAB, POS) m/z: 416 [M+H]⁺.

Example 28N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-ethyl-3-methyl-4-isoxazolecarboxamide

Like the Example 27 process, 5-ethyl-3-methylisoxazole-4-carboxylic acid(930 mg, 6.0 mmol) was made to react withN-(2-aminophenyl)-4-methoxybenzene sulfon amide (1.39 g, 5.0 mmol) toyield 1.96 g of the titled compound.

NMR (DMSO-d6) ppm: 1.28 (3H, t, J=7.6 Hz), 2.43 (3H, s), 3.04 (2H, q,J=7.6 Hz), 3.83 (3H, s), 6.80 (1H, dd, J=1.5, 7.9 Hz), 6.95-7.08 (3H,m), 7.21 (1H, dt, J=1.5, 7.9 Hz), 7.60 (2H, d, J=8.8 Hz), 7.89 (1H, dd,J=1.3, 8.1 Hz), 9.10 (1H, br s), 9.50 (1H, br s).

MS (FAB, POS) m/z: 416 [M+H]⁺.

Example 29N-(2-(4-methoxybenzenesulfonamide)phenyl]-3,5-diethyl-4-isoxazolecarboxamide

Like the Example 27 process, 3,5-diethyl-isoxazole-4-carboxylic acid(1.01 g, 6.0 mmol) was made to react withN-(2-aminophenyl)-4-methoxybenzene sulfon amide (1.39 g, 5.0 mmol) toyield 1.82 g of the titled compound.

NMR (DMSO-d6) ppm: 1.25 (3H, t, J=7.7 Hz), 1.29 (3H, t, J=7.7 Hz), 2.85(2H, q, J=7.7 Hz), 3.03 (2H, q, J=7.7 Hz), 3.83 (3H, s), 6.82 (1H, dd,J=1.5, 7.9 Hz), 6.96-7.10 (3H, m), 7.22 (1H, dt, J=1.5, 7.9 Hz), 7.60(2H, d, J=8.8 Hz), 7.88 (1H, dd, J=1.3, 8.1 Hz), 9.18 (1H, br s), 9.48(1H, br s).

MS (FAB, POS) m/z: 430 [M+H]⁺.

Example 30N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-isopropyl-3-methyl-4-isoxazolecarboxamide

Like the Example 27 process, 5-isopropyl-3-methylisoxazole-4-carboxylicacid (1.01 g, 6.0 mmol) was made to react withN-(2-aminophenyl)-4-methoxybenzene sulfon amide (1.39 g, 5.0 mmol) toyield 1.59 g of the titled compound.

NMR (DMSO-d6) ppm: 1.31, 1.34 (2×3H, each s), 2.43 (3H, s), 3.61 (1H,m), 3.85 (3H, s), 6.78 (1H, dd, J=1.5, 7.9 Hz), 6.96-7.08 (3H, m), 7.22(1H, dt, J=1.5, 7.9 Hz), 7.61 (2H, d, J=9.0 Hz), 7.92 (1H, dd, J=1.3,8.1 Hz), 9.12 (1H, br s), 9.50 (1H, br s).

MS (FAB, POS) m/z: 430 [M+H]⁺.

Example 31N-[2-(4-methoxybenzenesulfonamide)phenyl]-5-methyl-3-phenyl-4-isoxazolecarboxamide

Like the Example 27 process, 5-methyl-3-phenylisoxazole-4-carboxylicacid (500 mg, 2.46 mmol) was made to react withN-(2-aminophenyl)-4-methoxybenzene sulfonamide (570 mg, 2.05 mmol) toyield 400 mg of the titled compound.

NMR (DMSO-d6) ppm: 2.41 (3H, s), 3.80 (3H, s), 6.90-7.24 (5H, m),7.53-7.61 (5H, m), 7.73-7.88 (3H, m), 9.33 (1H, br s), 9.70 (1H, br s).

MS (FAB, POS) m/z: 464 [M+H]⁺.

Example 32N-[2-(4-methoxybenzenesulfonamide)phenyl]-3-methyl-5-phenyl-4-isoxazolecarboxamide

Like the Example 27 process, 3-methyl-5-phenylisoxazole-4-carboxylicacid (1.22 g, 6.0 mmol) was made to react withN-(2-aminophenyl)-4-methoxybenzene sulfonamide (1.39 g, 5.0 mmol) toyield 1.12 g of the titled compound.

NMR (DMSO-d6) ppm: 2.65 (3H, s), 3.80 (3H, s), 6.87-7.22 (5H, m),7.46-7.61 (5H, m), 7.68-7.78 (3H, m), 9.34 (1H, br s), 9.54 (1H, br s).

MS (FAB, POS) m/z: 464 [M+H]⁺.

The pharmacological effects of the compounds of the present inventionwill be demonstrated concretely byway of test examples. Abbreviationsused in the test examples are as follows,

CTP: Cytidine 5′-triphosphate

EGTA: Ethylene glycol bis(2-aminoethylether)

GTP: Guanosine 5′-triphosphate

MES: 2-morpholinoethane sulfonic acid

RB: Reaction buffer

Test Example 1

Tubulin polymerization inhibitory test using microtubule proteinsoriginated from swine brain: Microtubule proteins was extracted fromswine brain by the Shelansky method (Tanpakusitu jikkenhou,Zoku-Seikagaku Jikken Kouza, Vol.6, “Structure and Function ofCytoskeleton” (1^(st) Vol.),Tokyo Kagaku Dojin). Polymerization ofmicrotubule proteins was assayed by turbidity measurement.

Warming in RB (MES: 100 mM, MgCl₂: 0.5 mM, EGTA: 1 mM, pH6.8) containingGTP accelerates microtubule proteins, which are previously depolymerizedin ice bath, to reconstruct microtubules, with increased turbidityresulted in change in turbidity can be measured by an absorptiometer.The compounds of the present invention were dissolved indimethylsulfoxide to prepare their test solutions. To 245 μl of a RBsolution containing microtubule proteins (2 mg/ml) and GTP (1 mM) wasadded 5 μl of either an above test solution or dimethylsulfoxide toprepare a sample, which was incubated at 37° C. for 30 min. Absorbanceat 340 nm was measured by an absorptiometer, and Inhibitory Rate wascalculated by the equation as described below to determine IC₅₀, theconcentration of a test compound required to achieve 50% of inhibitoryrate. Absorbance at 340 nm of the compound was previously measured.

Inhibitory Rate(%)=[1−(T−C _(min))/(C _(max) −C _(min))]×100

where T is absorbance of the incubated sample with a test compound,C_(max) is absorbance of the incubated sample without the compound, andC_(min) is the non-incubated sample without the compound. The IC₅₀values are shown in the Table 1.

TABLE 1 Test compound IC₅₀ Test compound IC₅₀ (EXAMPLE No.) (μg/ml)(EXAMPLE No.) (μg/ml) 1 1.2 15 1.85 2 36 21 4.7 3 38 22 1.9 4 1.4 231.20 6 39 24 1.15 7 5.1 25 1.55 8 230 26 1.90 9 150 27 1.85 11 150 281.30 13 1.4 29 3.95 14 1.65 30 3.30 (note): IC₅₀ of colchicine (made byWako Junyaku, Tokyo) is 4.0 μg/ml.

Test Example 2

Anti-tumor in vitro test using the A2780 (humane ovarian cancer cell):

A2780 cells that were suspended on a RPMI 1640 culture medium containing10% calf serum, penicillin (50U/ml), and streptomycin (50 μg/ml) wereseeded in a 96 well flat microplate by 1000 cells (0.2 ml)/well, andcultured in a 5% carbon dioxide atmospheric incubator at 37° C. for 1day. The compounds of the present invention that were dissolved indimetylsulfoxide and diluted with a RPMI 1640 containing 10% calf serumto prepare the samples, provided that dimetylsulfoxide was restricted to0.2% and below in concentration.

The Supernatant liquids of the above A2780 cells wells were removed byaspiration, and then each 0.2 ml of the prepared samples were added tothe wells, which were incubated in a 5% carbon dioxide atmosphericincubator at 37° C. for 3 days. After the incubation, each 0.01 ml ofMTT solutions (3-(4,5-dimethylthiazole-2-yl)-2,5-dipheny-ltetra zoliumbromide, 5 mg/ml) were added to the wells, which were incubated foranother 2 hrs. After the supernatant liquids in the wells were removedby aspiration, the formazans formed in the wells were dissolved in 0.1ml of dimetylsulfoxide, and absorbances at 540 nm were measured by amicro plate reader to determine indices of live cells counts in thewells. Inhibitory Rate was calculated by the equation as described belowto determine IC₅₀, the concentration of a test compound required toachieve 50% of inhibitory rate.

Inhibitory Rate(%)=[(C−T)/C]×100

where T is absorbance in the well with a test compound and C isabsorbance in the well without the compound. The IC₅₀ values are shownin the Table 2.

TABLE 2 Test compound IC₅₀ Test compound IC₅₀ (EXAMPLE No.) (μg/ml)(EXAMPLE No.) (μg/ml) 1 0.023 17 1.4 2 0.58 20 6.8 3 0.22 21 0.079 40.017 22 0.14 5 7.2 23 0.006 6 0.14 24 0.006 7 0.20 25 0.05 8 6.7 260.034 9 13 27 0.034 11 0.68 28 0.013 12 9.7 29 0.14 13 0.010 30 0.16 140.036 31 0.30 15 0.025 32 0.62

Test Example 3

Prophylactic experiment of collagen-induced arthritis in mice:

The suspension of M. tuberculosis H37RA in Freund incomplete adjuvant (2mg/ml) was mixed with 0.3% type II collagen (extracted from bovinearticular cartilage, Collagen Gijutsu Kensyu Kai) in their equal amountsto form an emulsion, 0.1 ml of which was inected subcutaneously into thebase of the tail of mice to sensitize it. After twenty-one days, 0.3%type II collagen was th physiological saline solution to the one sixthconcentration of the solution, 0.2 ml of which was injectedintraperitoneally to sensitize secondarily. The mice that the firstsensitizations caused to fall into arthritis were excluded from thetest. The suspensions of the compounds (in the Example 13, 27, and 28)in 0.5% sodium carboxy methylcellulose (CMC-Na) were administratedorally to the test group mice once a day for 5 weeks from the day afterthe second sensitization. 0.5% CMC-Na solutions were administrated tothe control group mice to be compared with. Each group was composed ofeight mice. Daily symptoms in the mice limbs were observed to determinestratified scores (0 to 4, hence maximum 16). Scores and their standardsare as follows.

0: No symptom

1: Tumefaction or rubor in only one finger (or heel)

2: Tumefaction or rubor in two or above fingers (or heels) or in partsof fore (or hind) paw(s)

3: Tumefaction or rubor in the wholes of fore (or hind) paw(s) includingjoints

4: Ankylosis in the joints of finger(s) (or heel(s)) or fore (or hind)paw(s)

The results on the last day of this test are shown in the Table 3.

TABLE 3 Test compound Dose (EXAMPLE No.) mg/kg Averaged score Control 08.5 ± 1.7  13 25 2.3 ± 0.8* 13 50  0.4 ± 0.4** 27 50 1.7 ± 1.1* 28 503.0 ± 1.1* **p<0.005, *p<0.05

Compared with the control group, the test groups of the compounds of thepresent invention show significant prophylactic effect oncollagen-induceded arthritis model in mice which is an animal model forrheumatoid arthritis.

Furthermore, blood tests (blood cell count, GOT) and histologicalobservations of main organs executed on the last day that the compoundsgave little toxicity on bone marrow, liver heart, lung, spleen,pancreas, and gastrointestine.

INDUSTRIAL APPLICABILITY

The compounds of the present invention are confirmed to have tubulinpolyymerization inhibitory activity and anti-tumor effect. Theprophylactic experiment of collagen-induced arthritis in mice which isanimal model for rheumatoid arthritis confirms them to have excellentanti-rheumatic effect. Furthermore, they have low toxicity and 5 weeks'continuous administrations as executed in the Test Example give nodeath. Blood tests and histological observations suggest that thecompounds give little toxicity on bone marrow, liver, heart, lung,spleen, pancreas, and gastrointestine. Therefor, the compounds of thepresent invention are useful as low toxic anti-tumor agents andpreventives or remedies for rheumatism.

What is claimed is:
 1. A method for inhibiting tubulin polymerizationcomprising administering to an individual in need thereof an effectiveamount of the sulfonamide derivatives represented by the general formula(1):

wherein R¹ and R² may be the same or different and each independentlyrepresents a hydrogen atom, a halogen atom, a lower alkyl group, a loweralkoxy group, a nitro group, a hydroxy group; each R³, R⁴, R⁵ and R⁶represents a hydrogen atom; A represents any group of (1) 5-memberedheterocyclic group which is optionally substituted by a lower alkylgroup or phenyl group, whose ring members include at least 1 nitrogenatom and may include any atom(s) selected from the group consisting ofnitrogen atom, oxygen atom, and sulfur atom, (2) an alicyclic groupwhich is optionally substituted by a lower alkyl group or phenyl group,and (3) an alicyclic group whose ring members include at least 1nitrogen atom and may include any atom(s) selected from the groupconsisting of nitrogen atom, oxygen atom, and sulfur atom, orpharmaceutically acceptable salts thereof together with apharmaceutically acceptable excipient or carrier.
 2. A method fortreating rheumatism comprising administering to an individual in needthereof an effective amount of the sulfonamide derivatives as defined inclaim 1, or pharmaceutically acceptable salts thereof, together with apharmaceutically acceptable excipient or carrier.
 3. The method fortreating rheumatism according to claim 2, wherein R¹ is a lower alkoxygroup, and R² represents a hydrogen atom.
 4. The method for treatingrheumatism according to claim 3, wherein A represents an optionallysubstituted 5-membered heterocyclic group whose ring members include atleast 1 nitrogen atom and may include any atom(s) selected from thegroup consisting of nitrogen atom, oxygen atom, and sulfur atom.
 5. Themethod for treating rheumatism according to claim 4, wherein Arepresents an optionally substituted isoxazolyl group.
 6. The method fortreating rheumatism according to claim 5, wherein A represents a4-isoxazolyl group which has at least one lower alkyl group at the 3-and 5-position.
 7. The method for treating rheumatism according to claim6, wherein R¹ is a lower alkoxy group which is located at thepara-position.
 8. The method for treating rheumatism according to any ofclaims 2, 3, 4, 5, 6 or 7, wherein said rheumatism is inflammatoryrheumatism.
 9. A method for inhibiting tubulin polymerization accordingto claim 1, wherein R¹ is a lower alkoxy group, and R² represents ahydrogen atom.
 10. A method for inhibiting tubulin polymerizationaccording to claim 1, wherein A represents an optionally substituted5-membered heterocyclic group whose ring members include at least 1nitrogen atom and may include any atom(s) selected from the groupconsisting of nitrogen atom, oxygen atom, and sulfur atom.
 11. A methodfor inhibiting tubulin polymerization according to claim 10, wherein Arepresents an optionally substituted isoxazolyl group.
 12. A method forinhibiting tubulin polymerization according to claim 11, wherein Arepresents a 4-isoxazolyl group which has at least one lower alkyl groupat the 3- and 5-position.
 13. A method for inhibiting tubulinpolymerization according to claim 12, wherein R¹ is a lower alkoxy groupwhich is located at the para-position.